Methods to Test the Role of Age-related Lifestyle and Vaginal Microenvironment Changes and the Prevention, Treatment, and Progression of Genitourinary Syndrome of Menopause

超过50%的绝经后妇女受到绝经后生殖道综合征(GSM)的影响， 与雌激素下降有关的进行性慢性疾病，包括阴道萎缩，阴道干燥， 性欲降低，以及其他体征和症状。GSM症状是一个代价高昂的公共卫生问题， 如果不治疗，情况会恶化。更年期雌激素水平下降被认为会导致阴道蓄积减少 糖原和阴道微生物区系的变化。这些细菌包括低位阴道乳杆菌。50%-80%的 绝经后的女性。乳杆菌属部分通过乳酸保护泌尿生殖道免受病原体侵袭 产生和假想的抗炎特性，但乳杆菌是否在 GSM是未知的。现有的治疗外阴阴道GSM症状的方法都有局限性。有些女人是 激素治疗禁忌或担心副作用。虽然阴道润滑剂提供了 一些缓解，许多对阴道上皮有毒，减少乳杆菌，并增加泌尿生殖系统感染的风险。 因此，需要新的方法来治疗和预防GSM。因为乳酸菌在更年期时通常会减少， 阴道微生物区系是看似合理的治疗靶点；然而，益生菌本身并未被证明有效。 此外，与年龄相关的阴道微生物区系的变化与代谢物和免疫谱共同变化；但是，交叉- 这些分子特征和它们在GSM中的作用之间的对话尚不清楚。我们假设核心阴道 微环境生物标志物(VMB；例如微生物、代谢物和免疫图谱)反映阴道生物 衰老(V-BA)，在更年期增加，但可能会因生活方式因素而改变。前瞻性研究包括 需要确定与年龄相关的核心VMB，并确定它们如何影响GSM。为了实现这一目标，我们将 利用从812名年龄在35-60岁之间的临床女性收集的2,301份存档宫颈阴道样本 每六个月访问一次，为期两年(R01-CA123467)。具体目标是1)使用VMB对V-BA进行量化；2) 评价V-BA与GSM的纵向关系；3)纵向评价VBA与GSM的关系 生活方式因素对V-BA和VMB的影响；4)量化V-BA和VMB在生活方式和VMB之间的纵向中介作用 GSM。微生物区系已经通过16S rRNA基因扩增子序列(R21-AI107224)进行了分析。我们会 量化70种免疫标志物(细胞因子、趋化因子、生长因子)和代谢物的浓度 (GC/LC-MS)。这个提议的项目的一个新奇之处在于，我们将开发、验证和 对复杂的纵向数据应用新的统计方法，以改进和适应现代结构建模 和成分微生物组数据分析。这些方法将提供一个严格的框架来处理 数据缺失和混淆一直是许多微生物组研究的主要限制。终极公众 对健康的影响是，这个项目的发现有可能为开发新的治疗方法提供信息，以预防 或治疗GSM以促进女性健康。我们的团队在生物统计学、流行病学、老龄化、 这一系统所需的妇科学、基因组学、微生物学和免疫学--GSM的生物学方法。

Over 50% of postmenopausal women are affected by the genitourinary syndrome of menopause (GSM), a progressive, chronic condition related to estrogen decline that includes vaginal atrophy, vaginal dryness, reduced sexual desire, and other signs and symptoms. GSM symptoms are a costly public health concern and worsen if untreated. Estrogen decline in menopause is thought to lead to reduced accumulation of vaginal glycogen and shifts in vaginal microbiota. These include low vaginal Lactobacillus spp. levels in 50-80% of postmenopausal women. Lactobacillus spp. protect the urogenital tract from pathogens in part by lactic acid production and hypothesized anti-inflammatory properties, but whether lactobacilli play a functional role in GSM is unknown. Available treatments for vulvovaginal GSM symptoms have limitations. Some women are contraindicated for hormonal therapy or are concerned about side effects. While vaginal lubricants provide some relief, many are toxic to the vaginal epithelium, reduce lactobacilli, and raise urogenital infection risk. Thus, new ways to treat and prevent GSM are needed. Because lactobacilli are often reduced in menopause, vaginal microbiota are plausible treatment targets; however, probiotics alone have not been proven effective. Also, age-related changes in vaginal microbiota co-vary with metabolite and immune profiles; but, the cross- talk between these molecular features and their role in GSM is unknown. We hypothesize that core vaginal micro-environment biomarkers (VMB; e.g., microbial, metabolite, and immune profiles) reflect vaginal biological aging (V-BA) that increases in menopause but may be modifiable by lifestyle factors. Prospective studies are needed to identify core age-related VMB and determine how they affect GSM. To achieve this goal, we will leverage 2,301 archived cervicovaginal samples collected from 812 women aged 35-60 years with clinical visits every six months for two years (R01-CA123467). Specific Aims are to 1) quantify V-BA using VMB; 2) evaluate the longitudinal relationship between V-BA and GSM; 3) longitudinally assess the relationship of lifestyle factors on V-BA and VMB; 4) quantify longitudinal mediation by V-BA and VMB between lifestyle and GSM. Microbiota are already profiled by 16S rRNA gene amplicon sequencing (R21-AI107224). We will quantify concentrations of 70 immune markers (cytokines, chemokines, growth factors) and metabolites (GC/LC-MS) in cervicovaginal samples. A novelty of this proposed project is that we will develop, validate, and apply new statistical methods for complex longitudinal data that refine and adapt modern structural modeling and compositional microbiome data analyses. These methods will provide a rigorous framework to handle missing data and confounding that has been a major limitation in many microbiome studies. The ultimate public health impact is that findings from this project have potential to inform development of new therapy to prevent or treat GSM to promote women’s health. Our team has the expertise in biostatistics, epidemiology, aging, gynecology, genomics, microbiology, and immunology required for this systems-biology approach to GSM.